POLLEN MORPHOLOGY OF THALICTRUM L., SPECIES (RANUNCULACEAE) IN TURKEY

Pak. J. Bot., 37(2): 203-212, 2005. POLLEN MORPHOLOGY OF THALICTRUM L., SPECIES (RANUNCULACEAE) IN TURKEY SEVCAN TATLIDIL *, ADEM BICAKCI, HULUSI MALYER AND K.H. CAN BASER ** Department of Biology, Science Faculty, Uludag University, Gorukle, Bursa, Turkey ** Faculty of Pharmacy, Anadolu University, Eskisehir, Turkey Abstract In this study, the pollen morphology of 11 taxa of the Thalictrum L. (Ranunculaceae) viz., T. orientale Boiss., T. aquilegifolium L., T. sultanabadense Stapf.,T. foetidum L., T. isophyroides C.A.Mey., T.minus L., var. minus Boiss., T.minus var. majus (Crantz) Crepin, T. minus L. var. microphyllum Boiss., T.lucidum L., T. flavum L. and T. simplex L., from Turkey was investigated with light microscope and scanning electron microscope. LM observations reveal that pollen grains of the taxa are periporate with operculum, spheroidal, ornamentation microechinate. The numbers of pores are between 6 and 17. This palynological investigation is the first one which has been done on Thalictrum L., distributed in Turkey. Introduction The genus Thalictrum L., of the family Ranunculaceae comprises of approximately 150 species in the world (Hickey and King, 1965) of which and 11 taxa of the genus are found in Turkey (Davis, 1965). The genus Thalictrum L., has shown the highest complexity. Data on the pollen morphology of individual species of Thalictrum were obtained primarly by light microscopy (Kumazawa, 1936; Erdtman, 1943,1952; Erdtman et al., 1961; Kapp 1969; Petrow and Borrissova - Ivonova, 1975; Faegri and Iversen, 1992). Nilsson et al., (1977) studied pollen morphology of Thalictrum flavum L. using SEM. Moore and Webb (1978) have published a key including Thalictrum pollen. Pollen morphology of some Thalictrum species was studied by Punt et al., (1991) using LM and SEM. The present study aims to survey the pollen morphology of the Turkish representatives of Thalictrum using light and scanning electron microscopes and to discuss its significance in the taxonomy of the genus. The present study has been undertaken in order to provide further information which prove helpful in elucidating the taxonomy of the genus. Materials and Methods All the pollen samples were taken from herbarium specimens. Fig. 1 shows the distribution map of Thalictrum L., species in Turkey. For light microscopy, pollen grains were prepared by Erdtman s KOH-acetolysis technique; unstained residues were mounted in glycerin jelly (Erdtman, 1952). A Zeiss microscope with an apochromate oil immersion objective and periplan eyepiece was used for measuring the pollen diameter, exine thickness, pores shapes and mesoporium (Table 1). Measurements have been made with apiece micrometer (interval between two marks 0.62µm). At least 50 grains of each species were measured. Measure means (M) and standard deviation (S) were calculated. For this purpose the Excel 4.0 program for Windows was used. * Corresponding Author: sevcant@uludag.edu.tr

204 SEVCAN TATLIDIL ET AL., Fig. 1. Distribution map of Thalictrum L., species in Turkey. 1- T. orientale; 2- T. foetidum; 3- T. minus var. majus; 4- T. flavum; 5- T. aquilegifolium; 6- T. isophyroides; 7- T. minus var. microphyllum; 8- T. simplex; 9- T. sultanabadense; 10- T. minus var. minus; 11- T. lucidum.

POLLEN MORPHOLOGY OF THALICTRUM SPECIES IN TURKEY 205

206 SEVCAN TATLIDIL ET AL., All taxa were examined using SEM except T. sultanabadense Stapf., for which polliniferous material was not enough. For scanning electron microscopy, the pollen grains were transferred directly to double-sided tape affixed stub and vacuum coated with gold. Photomicrographs were taken using JEOL 540 scanning electron microscope in Anadolu University, Turkey. The descriptive terminology follows that of Faegri & Iversen (1992) and Punt et al., (1991). Results Description of pollen grains T. aquilegifolium L. (Table 1; Fig. 2, a-b) Pollen grains are pantoporate (6-9 pores), apolar, spheroidal, 22 µm in diameter.pore circular, 4.78 µm in diameter. Distance between pores 6.68 µm. Pore membrane with spaced varts and pores margin flattened Exine thickness 1.2 µm. Ectexine slightly thicker than or same thickness as endexine. Ornamentation micro-echinate T. flavum L. (Table 1; Fig. 2, c-d) Pollen grains are pantoporate (6-10 pores), apolar, spheroidal, 17.51-µm in diameter. Pores circular, 3.21µm in diameter. Distance between pores 6.65 µm. Pore membrane with spaced varts and pores margin flattened. Exine thickness 1.2µm Ectexine slightly thicker than or same thickness as endexine. Ornamentation micro-echinate T. foetidum L. (Table 1; Fig. 2, e-f) Pollen grains are pantoporate, (6) 8 10 (13 pores), apolar, spheroidal, 21.02 µm in diameter.pore circular, 3.42 µm in diameter. Distance between pores 6.45 µm. Pore membrane with densely spaced granules, pores margin flattened or sunken. Exine thickness 1.22 µm. Ectexine slightly thicker than or of same thickness as endexine. Ornamentation micro-echinate. T. isophyroides C.A. Mey. (Table 1; Fig. 3, a-b) Pollen grains are pantoporate (6-11 pores), apolar, spheroidal, 19.04 µm in diameter. Pore circular, 4.25 µm in diameter. Distance between pores 7.02 µm. Pore membrane with spaced varts and pores margin flattened or sunken. Exine thickness 1.2µm. Ectexine slightly thicker than or of same thickness as endexine. Ornamentation micro-echinate T. lucidum L. (Table 1; Fig. 4, g-h) Pollen grains are pantoporate (8-11 pores), apolar, spheroidal, 21.18 µm in diameter. Pore circular, 4.43 µm in diameter. Distance between pores 5.76 µm. Pore membrane with densely spaced granules and pores margin flattened. Exine thickness 1.2 µm. Ectexine slightly thicker than or of same thickness as endexine. Ornamentation microechinate

POLLEN MORPHOLOGY OF THALICTRUM SPECIES IN TURKEY 207 a b c d e f Fig. 2. Scanning electron micrographs of Thalictrum L.; T. aguilegifolium (a-b); T. flavum (c-d); T. foetidum (e-f); scale bar: a, c, e - 10µm; b, d, f - 5µm.

208 SEVCAN TATLIDIL ET AL., a b c d e f Fig. 3. Scanning electron micrographs of Thalictrum L.; T. İsophyroides (a-b) T. minus var. microphyllum (c-d); T. minus var.majus (e-f); scale bar: a, c, e) 10µm; b, d, f - 5µm.

POLLEN MORPHOLOGY OF THALICTRUM SPECIES IN TURKEY 209 T. minus L.var. minus Boiss. (Table 1; Fig. 4, a-b) Pollen grains are pantoporate (11-17 pores), apolar, spheroidal, 28.24 µm in diameter. Pore circular, 3.32 µm in diameter. Distance between pores 5.97 µm. Pore membrane with spaced spinules and pores margin sunken Exine thickness 1.2µm. Ectexine slightly thicker than or of same thickness as endexine. Ornamentation microechinate T.minus var. majus (Crantz) Crepin (Table 1; Fig. 3, e-f) Pollen grains are pantoporate (8-11 pores), apolar, spheroidal, 24.99 µm in diameter. Pores circular, 5.04 µm in diameter. Distance between pores 7.91 µm. Pore membrane with spaced gemma and pores margin flattened. Exine thickness 1.2µm. Ectexine slightly thicker than or of same thickness as endexine. Ornamentation micro-echinate T. minus var. microphyllum Boiss. (Table 1; Fig. 3, c-d) Pollen grains are pantoporate (6-12 pores), apolar, spheroidal, 20.21µm in diameter. Pores circular, 3.05 µm in diameter. Distance between pores 6.34 µm. Pore membrane with spaced spinules and pores margin flattened. Exine thickness 1.2µm. Ectexine slightly thicker than or of same thickness as endexine. Ornamentation micro-echinate T. orientale Boiss. (Table 1; Fig. 4, c-d) Pollen grains are pantoporate (6-13 pores), apolar, spheroidal, 21µm in diameter. Pore circular, 3.11µm in diameter. Distance between pores 5.02 µm. Pore membrane with densely spaced granules and pores margin flattened. Exine thickness 1.2µm. Ectexine slightly thicker than or same thickness as endexine. Ornamentation micro-echinate T. simplex L. (Table 1; Fig. 4, e-f) Pollen grains are pantoporate (9-15 pores), apolar, spheroidal, 19.37 µm in diameter. Pore circular, 3.64 µm in diameter. Distance between pores 6.2 µm. Pore membrane with spaced varts and pores margin flattened. Exine thickness 1.2µm. Ectexine slightly thicker than or of same thickness as endexine. Ornamentation micro-echinate. T. sultanabadense Stapf. (Table 1) Pollen grains are pantoporate (6-11 pores), apolar, spheroidal, 19.43 µm in diameter. Pore circular, 3.67 µm in diameter. Distance between pores 6.2 µm. Pore membrane with densely spaced granules. Exine thickness 1.2µm. Ornamentation micro-echinate. Discussion Ranunculaceae is an europalynous family. Pollen grains (2-) 3 (4) colpate, 3 colporoidate, pantocolpate, pantoporate, nonaperturate; pollen shape oblat to spherpidae; ornamentation shows variety (echinate, reticulate etc.) (Erdtman, 1952).

210 SEVCAN TATLIDIL ET AL., a b c d e f g h Fig. 4. Scanning electron micrographs of Thalictrum L.; T. minus var. minus (a-b); T. orientale (cd); T. simplex (e-f); T. lucidum L. (g-h); scale bar: a, c, e, g - 10µm; b, d, f, h - 5µm.

POLLEN MORPHOLOGY OF THALICTRUM SPECIES IN TURKEY 211 Few studies have been made on the pollen of some Thalictrum L., species. Faegri and Iversen (1992) have published a key including Thalictrum pollen and described as 10-25 µm in diameter and 4-12 pores. T. aquilegifolium L., was described by Erdtman (1952) having 6-10 pores and 15-24µm in diameter. In our study, pollen grain of T. aquilegifolium was 22.2 µm in diameter with 6-10 pores. During investigation on pollen morphology of Thalictrum flavum L., Erdtman (1943) described that pollen of T. flavum is 16 µm in diameter, 8 pores and reticulate ornamentation. Petrov et al., (1975) found pores number 20 in T. flavum Nilsson (1977) studied pollen morphology of T. flavum using SEM showing 15-20-24 µm in diameter, pores number 6-10, pores diameter 5-6 µm. Moore and Webb (1978) reported 9 or more pores in T. flavum. Punt et al., (1991) found that pollen diameter was 18-(20)-24 µm, pores number 8-(12)-15 in T. flavum. In our study pollen of T. flavum are spheroidal, 17.51x 17.26 µm in diameter, 6-10 pores, pores diameter 3.21 µm. The pollen morphology of T. flavum correspond more closely to that reported by Nilsson (1977). Other studies are different from our studies especially in pores number. Kumazawa (1936) reported less than 10 pores in T. simplex L. In our study pores number of T simplex was found between 9 and 15. The most important information on the comparative pollen morphology of Thalictrum is given by Punt & Blackmore (1991). They have provided detailed palynomorphological description of six. The Northwest European species of Thalictrum. In the present study we found that, the pollen grains of 11 taxa of Thalictrum are pantoporate (6-17 pores), apolar, spheroidal. Pore membrane show different palynologic characters. Pores margin situated flattened or slightly sunken areas of the exine. Mezoporium 5.02-7.91 µm. Exine thickness 1.2µm, ectexine slightly thicker than or of same thickness as endexine. Ornamentation micro-echinate. Pollen morphological key to species of Thalictrum L., distributed in Turkey: 1.Operculum densely spaced granules or spinules 2. Pores margin flattened 3. Some pores margin flattened or some of them sunken... T. foetidum 3. All of pores margin flattened 4. Largest diameter of grains > 20 µm.... T. sultanabadense 4. Largest diameter of grains < 20 µm 5. Mesoporium > 6 µm....... T. orientale 5. Mesoporium < 6 µm...... T. minus var.microphyllum 2. Pores margin flattened 6. Operculum spaced granules..... T. lucidum 6. Operculum spaced spinules... T. minus var. minus 1. Operculum densely spaced varts or gemmae 7. Pores margin flattened 8. Some pores margin flattened or some of them sunken... T. isophyroides 8. All of pores margin flattened 9. Operculum spaced gemmae.. T. minus var. majus 9. Operculum spaced varts 10. Largest diameter of grains > 20 µm.... T. flavum 10. Largest diameter of grains < 20 µm T. aquilegifolium 7. Pores margin sunken.... T. simplex

212 SEVCAN TATLIDIL ET AL., In the flora of Turkey, it is described that T. simplex L. and T. flavum are related to each other (Davis, 1965). We have found that these two species are different from each other in the terms of palynologic characters. According to Davis (1965), T. minus L., is complex species for Turkey. In our study, pollen grains of T. minus L., var. minus are different from other species it is 24 µm in diameter and pores are between 11 and 17. This palynological investigation is the first one which has been done on Thalictrum species distributed in Turkey. References Davis, P.H.1965. Flora of Turkey and the East Aegean Islands. Volume I, Edinburgh University Press. Edinburgh. 567 pp. Erdtman, G.1943. An Introduction to Pollen Analysis. Academic Press, New York. 712 pp. Erdtman, G. 1952. Pollen Morphology and Plant Taxonomy. Angiosperms. Almquvist and Wiksell, Stockholm. 553 pp.. Erdtman, G., B. Berglund and J. Praglowski.1961. An Introduction to A Scandinavian Pollen Flora. Grana Palynol., 2(2): 3-92. Faegri, K. and J. Iversen. 1992. Textbook of Pollen Analysis, Alden Press. London. 370 pp. Hickey, M. and C.J. King. 1965. 100 Families of Flowering Plants, Cambridge University Press, USA. 199 pp. Kapp, R.O.1969. How to Know Pollen and Spores? W.M.C. Brown Company Press. America..190 pp. Kumazawa, M. 1936. Pollen Grain Morphology in Ranunculaceae, Lardizabalaceaae and Berberidaceae. Jap. J. Bot., 8: 19-46. Moore, P. and J. Webb. 1978. Pollen Analysis. Department of Plant Sciences, Kings College, Second Edition, London. 459 pp. Nilsson, S., J. Praplowski and L. Nilsson. 1977. Atlas of Airborne Pollen Grains and spores in Northern Europe. Printed in Sweden by Ljungföretagen, 159 pp. Petrov, S. and O. Borrissova-Ivanova.1975. Pollen morphology of the Bulgarian representatives of the family Ranunculaceae. Juss. II. Thalictrum L. In: Festschrift in honour of Acad. Daki Jordanov, Sofia, pp. 101-112. Punt, W. and S. Blackmore. 1991. The Northwest European Pollen Flora. Vol VI, Elsevier Science, New York, USA. 275 pp. (Received for publication 8 January 2004)